Abstract
Phenylalanine, tyrosine, glycine, cystine, and phosphoethanolamine are commonly measured amino acids in various physiological fluids to diagnose or follow-up various inborn errors of metabolism. The gold standard method for the amino acids quantitation has been ion exchange chromatography with ninhydrin post-column derivatization. However, this method is very laborious and time consuming. In recent years, liquid-chromatography mass spectrometry is being increasingly used for the assay of amino acids. Pre-column butyl derivatization with reverse phase chromatography has been widely used for mass spectrometry analysis of amino acids. Phosphoethanolamine is not butylated and cannot be measured by this method. Nevertheless, phosphoethanolamine can be dansyl-derivatized using dansyl chloride. We developed a double derivatization method by using butanol and dansyl chloride to derivatize carboxylic and amino groups separately, and then combining the derivatives to simultaneously measure these five amino acids using TOF-MS scan. Stable isotope-labeled internal standards were used.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Blaskovics ME, Schaeffler GE, Hack S (1974) Phenylalaninaemia. Differential diagnosis. Arch Dis Child 49:835–843
Broer S, Palacin M (2011) The role of amino acid transporters in inherited and acquired diseases. Biochem J 436:193–211
Mattoo A, Goldfarb DS (2008) Cystinuria. Semin Nephrol 28:181–191
Dinopoulos A, Matsubara Y, Kure S (2005) Atypical variants of nonketotic hyperglycinemia. Mol Genet Metab 86:61–69
Imbard A, Alberti C, Armoogum-Boizeau P, Ottolenghi C, Josserand E, Rigal O, Benoist JF (2012) Phosphoethanolamine normal range in pediatric urines for hypophosphatasia screening. Clin Chem Lab Med 50:2231–2233
Mc CR, Morrison AB, Dent CE (1955) The excretion of phosphoethanolamine and hypophosphatasia. Lancet 268:131
Deyl Z, Hyanek J, Horakova M (1986) Profiling of amino acids in body fluids and tissues by means of liquid chromatography. J Chromatogr 379:177–250
Dietzen DJ, Weindel AL, Carayannopoulos MO, Landt M, Normansell ET, Reimschisel TE, Smith CH (2008) Rapid comprehensive amino acid analysis by liquid chromatography/tandem mass spectrometry: comparison to cation exchange with post-column ninhydrin detection. Rapid Commun Mass Spectrom 22:3481–3488
Hardy DT, Hall SK, Preece MA, Green A (2002) Quantitative determination of plasma phenylalanine and tyrosine by electrospray ionization tandem mass spectrometry. Ann Clin Biochem 39:73–75
Song C, Zhang S, Ji Z, Li Y, You J (2015) Accurate Determination of Amino Acids in Serum Samples by Liquid Chromatography-Tandem Mass Spectrometry Using a Stable Isotope Labeling Strategy. J Chromatogr Sci [in press]
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Deng, S., Scott, D., Garg, U. (2016). Quantification of Five Clinically Important Amino Acids by HPLC-Triple TOF™ 5600 Based on Pre-column Double Derivatization Method. In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Biomolecular Analysis. Methods in Molecular Biology, vol 1378. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3182-8_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3182-8_6
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3181-1
Online ISBN: 978-1-4939-3182-8
eBook Packages: Springer Protocols